US5288610A - Detecting reagent for antiplatelet antibody - Google Patents
Detecting reagent for antiplatelet antibody Download PDFInfo
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- US5288610A US5288610A US07/508,922 US50892290A US5288610A US 5288610 A US5288610 A US 5288610A US 50892290 A US50892290 A US 50892290A US 5288610 A US5288610 A US 5288610A
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- platelet
- carrier particle
- detecting reagent
- antibody
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- 230000000702 anti-platelet effect Effects 0.000 title claims abstract description 28
- 239000003146 anticoagulant agent Substances 0.000 title claims abstract description 28
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 17
- 239000002245 particle Substances 0.000 claims abstract description 31
- 239000000427 antigen Substances 0.000 claims abstract description 25
- 102000036639 antigens Human genes 0.000 claims abstract description 25
- 108091007433 antigens Proteins 0.000 claims abstract description 25
- 230000004520 agglutination Effects 0.000 claims abstract description 15
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- 238000006243 chemical reaction Methods 0.000 claims description 17
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- 239000006166 lysate Substances 0.000 claims description 6
- TUSDEZXZIZRFGC-UHFFFAOYSA-N 1-O-galloyl-3,6-(R)-HHDP-beta-D-glucose Natural products OC1C(O2)COC(=O)C3=CC(O)=C(O)C(O)=C3C3=C(O)C(O)=C(O)C=C3C(=O)OC1C(O)C2OC(=O)C1=CC(O)=C(O)C(O)=C1 TUSDEZXZIZRFGC-UHFFFAOYSA-N 0.000 claims description 5
- 239000001263 FEMA 3042 Substances 0.000 claims description 5
- LRBQNJMCXXYXIU-PPKXGCFTSA-N Penta-digallate-beta-D-glucose Natural products OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-PPKXGCFTSA-N 0.000 claims description 5
- 238000001179 sorption measurement Methods 0.000 claims description 5
- LRBQNJMCXXYXIU-NRMVVENXSA-N tannic acid Chemical compound OC1=C(O)C(O)=CC(C(=O)OC=2C(=C(O)C=C(C=2)C(=O)OC[C@@H]2[C@H]([C@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)[C@@H](OC(=O)C=3C=C(OC(=O)C=4C=C(O)C(O)=C(O)C=4)C(O)=C(O)C=3)O2)OC(=O)C=2C=C(OC(=O)C=3C=C(O)C(O)=C(O)C=3)C(O)=C(O)C=2)O)=C1 LRBQNJMCXXYXIU-NRMVVENXSA-N 0.000 claims description 5
- 229940033123 tannic acid Drugs 0.000 claims description 5
- 235000015523 tannic acid Nutrition 0.000 claims description 5
- 229920002258 tannic acid Polymers 0.000 claims description 5
- 108010010803 Gelatin Proteins 0.000 claims description 3
- 229920000159 gelatin Polymers 0.000 claims description 3
- 239000008273 gelatin Substances 0.000 claims description 3
- 235000019322 gelatine Nutrition 0.000 claims description 3
- 235000011852 gelatine desserts Nutrition 0.000 claims description 3
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- 239000002502 liposome Substances 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 241001465754 Metazoa Species 0.000 claims description 2
- 230000000961 alloantigen Effects 0.000 claims 4
- 210000003743 erythrocyte Anatomy 0.000 claims 3
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- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 9
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- WHTVZRBIWZFKQO-AWEZNQCLSA-N (S)-chloroquine Chemical compound ClC1=CC=C2C(N[C@@H](C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-AWEZNQCLSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 229960003677 chloroquine Drugs 0.000 description 3
- WHTVZRBIWZFKQO-UHFFFAOYSA-N chloroquine Natural products ClC1=CC=C2C(NC(C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-UHFFFAOYSA-N 0.000 description 3
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- 206010021245 Idiopathic thrombocytopenic purpura Diseases 0.000 description 1
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- 206010043561 Thrombocytopenic purpura Diseases 0.000 description 1
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- 208000007502 anemia Diseases 0.000 description 1
- 230000002391 anti-complement effect Effects 0.000 description 1
- 108010008730 anticomplement Proteins 0.000 description 1
- 201000003710 autoimmune thrombocytopenic purpura Diseases 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 150000001718 carbodiimides Chemical class 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- QSWDMMVNRMROPK-UHFFFAOYSA-K chromium(3+) trichloride Chemical compound [Cl-].[Cl-].[Cl-].[Cr+3] QSWDMMVNRMROPK-UHFFFAOYSA-K 0.000 description 1
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- 230000008588 hemolysis Effects 0.000 description 1
- 230000003100 immobilizing effect Effects 0.000 description 1
- 210000003593 megakaryocyte Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
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- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
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- 206010043554 thrombocytopenia Diseases 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56966—Animal cells
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S435/00—Chemistry: molecular biology and microbiology
- Y10S435/961—Chemistry: molecular biology and microbiology including a step of forming, releasing, or exposing the antigen or forming the hapten-immunogenic carrier complex or the antigen per se
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/829—Liposomes, e.g. encapsulation
Definitions
- the present invention relates to a detecting reagent for an antiplatelet antibody.
- This detecting reagent is used to detect an antiplatelet antibody present in a blood of a patient at the time of, e.g., platelet transfusion.
- Platelets are akaryocites which are present in a blood and each of which has a diameter of 2 to 3 ⁇ m.
- the platelets are produced by fragmentation of megakaryocytes present in the bone marrow.
- When platelet in a blood are brought into contact with damaged hemangioendothelial cells, they agglutinate with each other and play an important role for hemostasis.
- a platelet transfusion therapy has been made to aim at hemostasis and hemorrhagic prevention for patients who have a hemorrhagic tendency caused by thrombocytopenia or thrombopathy.
- This therapy many lot of patients are saved from death caused by hemorrhage, and their lives can be saved or prolonged.
- the amount of a platelet preparation used has been abruptly increased, with development of platelet transfusion.
- An antibody against the antigen present on a surface membrane of the transfused platelet is produced in case that a large amount of platelets must be frequently transfused over a long period of time.
- Examples of such case is the platelet transfusion therapy for patients who suffer from acute leukemia and asplastic anemia. When this occurs, platelets transfused from random donors cannot cause any desired response of the patients, and no increase in the number of platelets is found after platelet transfusion. When platelet transfusion does not function well due to the reason described above, platelets having an antigen type which does not react with an antiplatelet antibody of such a patient must be transfused to this patient.
- Antiplatelet antibodies are classified into an alloantibody and an autoantibody.
- the antiplatelet alloantibody is produced by the above-mentioned platelet transfusion or pregnancy.
- the isoantibody does not only influence an effect of platelet transfusion described above but also causes post-transfusion purpura and neonatal thrombocytopenic purpura.
- the antiplatelet autoantibody is produced in a patient who suffers from an autoimmune disease such as idiopathic thrombocytopenic purpura.
- the autoantibody is deemed to be a substance which causes an autoimmune disease.
- This method is the first method developed to detect the antiplatelet antibody and is originally used to detect a P1 A antigen series and a Ko antigen series. According to this method, an IgM antibody is mainly detected, but an IgG antibody is rarely detected. This method tends to cause nonspecific agglutination caused by natural agglutination of platelets and has a low sensitivity.
- hemolysin labeled sheep blood cell After a sample serum, platelets, and a complement are reacted with each other, hemolysin labeled sheep blood cell are added to the reaction mixture, and an amount of consumption of the complement is determined from a degree of hemolysis of the hemolysin labeled sheep blood cell.
- This method can detect an HLA antigen on a surface membrane of the platelet and a blood group antigen of ABO type, but has a low sensitivity. In addition, it is difficult to detect the antiplatelet antibody if a sample serum has anticomplement activity.
- the reaction mixture After platelets are reacted with an inactivated sample serum, the reaction mixture is washed. After an antiglobulin serum is added to the mixture, the mixture is subjected to a centrifugal operation. An anti-D antibody labeled blood cell is added to and reacted with the supernatant of the reaction mixture to detect an antiplatelet antibody in accordance with an antiglobulin titer of the supernatant. This method tends to cause pseudopositive determination and has a low sensitivity.
- the RIA is an antiglobulin test using an anti-human IgG labeled with a radioactive isotope.
- the isotope-labeled anti-human IgG is coupled to an antiplatelet antibody of a platelet of a patient, and the radioactive isotope elements coupled to the platelets are counted.
- Platelets of solid phase are formed on the well wall of a U-shaped microplate, and a sample serum is added to and reacted with the platelets. After the well is washed, anti-human IgG labeled sheep red cell is added to the reaction mixture and is reacted with it. An agglutination pattern in which the sheep red cells collect in a central portion of the well is determined to be negative, and an agglutination pattern in which sheep red cells are scattered on the entire area is determined to be positive, thereby detecting a platelet antibody.
- the agglutination assay (1) requires only one step for the entire reaction and has simple test procedures.
- the agglutination assay however, has disadvantages in that the IgG class antibodies cannot be detected and nonspecific agglutination tends to occur. Therefore, the agglutination assay is not suitable for practical applications.
- the present invention has been made in consideration of the above problems, and has as its object to provide a detecting reagent for an antiplatelet antibody, which can detect an antiplatelet antibody with a passive haemagglutination reaction in one step.
- a detecting reagent for an antiplatelet antibody comprising:
- a platelet antigen component immobilized on a surface of the carrier particle a platelet antigen component immobilized on a surface of the carrier particle.
- the present inventor made extensive studies to develop a simple method of measuring an antiplatelet antibody within a short period of time and was successful in detection of an antiplatelet antibody in a sample serum by a passive haemagglutination reaction in one step upon mixing of the sample serum with a carrier particle on which an antigen component of the platelet is immobilized.
- the antigen component of the platelet can be obtained as a solubilized lysate upon application of a physical force such as an ultrasonic force to a platelet.
- the antigen component ca also be obtained by solubilizing the platelet upon a treatment of the platelet with a surfactant.
- a physical force is applied to the platelet to crush it, and the crushed platelet pieces are treated with a surfactant to solubilize the platelet, thereby obtaining the antigen component.
- the carrier particle used in the present invention may be any particle which can be used in the passive agglutination reaction.
- the carrier particle are a red cell of man or an animal (e.g, sheep or chicken), a liposome, a latex particle, and a gelatin particle.
- a known method can be used as a method of immobilizing an antigen component of a platelet on a carrier particle, and is exemplified by a method using a coupling agent such as tannic acid, glutaraldehyde, carbodiimide, or chromium chloride.
- a coupling agent such as tannic acid, glutaraldehyde, carbodiimide, or chromium chloride.
- its coupling agent is N- ⁇ -(aminoethyl)- ⁇ -aminopropyl-trimethyoxysilane.
- the antigen component may be immobilized on the carrier particle by physical adsorption without using any coupling agent.
- particles of the reagent do not agglutinate with each other, contrary to the platelets subjected to natural agglutination, propably because the platelet membrane antigen component immobilized on the carrier particle is solubilized in advance. Therefore, a good measurement result with good reproducibility can be obtained without causing natural agglutination between the particles.
- an antiplatelet antibody can be detected by a passive haemagglutination reaction in one step.
- a human whole blood of type O was collected at a volume ratio of 7:1 with respect to an ACD-A solution (available from Terumo Corp.), and the whole blood added with the ACD-A solution was subjected to centrifugal separation at a speed of 1,400 rpm for 10 minutes, thereby separating a platelet-rich plasma (PRP).
- PRP platelet-rich plasma
- an ACD-A solution was added to the resultant PRP in an amount of 10% by volume of the PRP, and the resultant mixture was subjected to centrifugal separation at a speed of 2,500 rpm for 15 minutes. The supernatant was removed, and a platelet concentrate (P.C.) was obtained. This platelet concentrate was washed with a physiological saline, and finally a platelet suspension solution (about 1 ⁇ 10 5 platelets/ ⁇ l) was obtained.
- the resultant platelet suspension solution was dropped in an amount of 5 ⁇ l on each well of a styrol microplate (Terasaki Plate U Type (available from Robins Corp.)) electrostatically discharged in advance.
- the microplate was placed on a plate mixer to stir the solution in each well for 10 seconds.
- the microplate was then set in a plate centrifugal separator, and the solution in each well was centrifugally separated at 2,000 rpm for 5 minutes, so that the platelets were coated on to the bottom surface of each well.
- a fixing solution was prepared by diluting a 35% formalin solution to five times with a PBS (phosphate buffered saline), and adjusting a pH value to 7.2 with addition of NaOH.
- the fixing solution thus obtained was added to each well by 10 l/well and was left to stand for 20 minutes. After each well was washed with a physiological saline three times, it was confirmed with naked eyes that a single layer of platelets was formed on the bottom surface of each well. Preparation of typing microplates were thus completed.
- Antiserums used in Example 1 were human anti-serums (e.g., anti-P1 A1 anti-P1 A2 anti-Yuk a , anti-Yuk b , anti-Bak a , anti-Nak a , anti-Sib a , anti-Ko b , and anti-Br a ) against platelet-specific antigens, and an anti-serums (e.b., anti-HLA) against an HLA antigen.
- human anti-serums e.g., anti-P1 A1 anti-P1 A2 anti-Yuk a , anti-Yuk b , anti-Bak a , anti-Nak a , anti-Sib a , anti-Ko b , and anti-Br a
- an anti-serums e.b., anti-HLA
- the platelets of donor No. 1 which were positive for the anti-P1 A1 , anti-Yuk b , anti-Bak a , anti-Nak a , anti-Sib a , and anti-Ko a were solubilized by the following procedures.
- chloroquine 0.8% chloroquine was added to a platelet suspension solution (105/ ⁇ l) and was reacted with it at room temperature for an hour.
- the reacted platelets were washed with a physiological saline twice, and 2.5-ml platelets were crushed with sonic oscillation by a Sonica-tor (available from Tomy Corp.) for 20 seconds.
- 10 5 G of the crushed platelets were centrifugally separated for 30 minutes and washed once.
- 1% Triton X-100 was added to the separated platelets.
- the platelets were further centrifugally separated at 10 5 G for 30 minutes, and the platelet membrane fraction in the supernatant was sampled to obtain a solubilized platelet membrane antigen.
- the HLA antigen was removed by the chloroquine treatment.
- the chloroquine treatment however, can be omitted to cause both the platelet specific antigen and the HLA antigen to exist in the finally obtained platelet membrane antigen fraction.
- the PBS was added to a pellet of the washed red cells to prepare a 5% red cell suspension.
- 20 ml of 2.5% glutaraldehyde solution were added to the 100 ml of the red cell suspension and were shaken and reacted with it at room temperature for 3 hours.
- the sheep red cells to which a carbonyl group was introduced was washed with a PBS four times to add the PBS, thereby obtaining a 5% suspension.
- the platelet membrane antigen labeled sheep red cell (to be referred to as an indicator cell hereinafter) was used to perform an agglutination reaction with the antiplatelet antibody in accordance with the following procedures.
- a PBS pH: 7.2 was added to the first to third wells of a U-shaped microplate by 25 ⁇ l/well. 25 ⁇ l of a sample serum were poured in the first well and were stirred well (dilution: twice). Similarly, serum samples diluted four times and eight times were respectively dispensed into the second and third wells, respectively. Serum samples diluted 2 n times were sequentially dispensed in the fourth and subsequent wells. The indicator cells were added to each well by 25 ⁇ l/well, and were stirred well. The lid of each cell was then closed, and each mixture was left to stand at room temperature for 2 hours. Agglutination patterns formed on the bottoms of the wells were determined.
- Sample serums were a human serum of a healthy man, an anti-P1 A1 serum, an anti-Yuk b serum, an anti-Bak a serum, an anti-Nak a serum, an anti-Sib a serum, an anti-Ko a serum, and an anti-HLA serum.
- the reaction with the serum of the healthy man exhibited a pseudopositive up to a dilution ratio of 1:4, but were negative when the dilution ratio was 1:8 or more.
- the P1 A1 and anti-Yuk b antibodies could be detected with a sensitivity equivalent to that in the MPHA method.
- the Nak a antibody could be detected although the reaction was rather weak.
- anti-Bak a anti-Sib a
- anti-Ko a antibodies could not be detected.
- the indicator cell obtained in the above example reacts with the anti-P1 A1 anti-Yuk b , and anti-Nak a antibodies, but does not react with the anti-Bak a , anti-Sib a , and anti-Ko a antibodies. Therefore, the present invention is effective for identifying patients who possess antibodies against the anti-P1 A1 , anti-Yuk b , and anti-Nak a antibodies.
- the indicator cell of this example may be combined with an indicator cell having another specificity to effectively perform screening of patients who possess antiplatelet antibodies and typing of antiplatelet antibodies.
- solubilized platelet membrane antigen is used in the indicator cell of this example, nonspecific agglutination between the indicator cells does no occur, thereby achieving good reproducibility.
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Abstract
Description
TABLE 1 ______________________________________ Anti-Serum Donor Anti- Anti- Anti- Anti- Anti- No. Pl.sup.A1 Pl.sup.A2 Yuk.sup.a Yuk.sup.b Bak.sup.a ______________________________________ 1 + - - + + 2 + - - + + 3 + - - + + 4 + - - + - 5 - - - - - ______________________________________ Anti-Serum Donor Anti- Anti- Anti- Anti- Anti- No. Nak.sup.a Sib.sup.a Ko.sup.a Br.sup.a HLA ______________________________________ 1 + + + - - 2 + - - - - 3 - - - - - 4 - - - - - 5 - - - - - ______________________________________
Claims (9)
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1-122392 | 1989-05-16 | ||
JP1122392A JPH02300664A (en) | 1989-05-16 | 1989-05-16 | Reagent for detecting antiplatelet antibody |
Publications (1)
Publication Number | Publication Date |
---|---|
US5288610A true US5288610A (en) | 1994-02-22 |
Family
ID=14834661
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US07/508,922 Expired - Lifetime US5288610A (en) | 1989-05-16 | 1990-04-12 | Detecting reagent for antiplatelet antibody |
Country Status (2)
Country | Link |
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US (1) | US5288610A (en) |
JP (1) | JPH02300664A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0762121A1 (en) * | 1995-08-09 | 1997-03-12 | Nippon Zoki Pharmaceutical Co., Ltd. | Antigen for enzyme immunoassay and method of measuring anti-erythrocyte antibody |
Families Citing this family (1)
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EP0593572B1 (en) * | 1991-07-01 | 1998-06-17 | The Blood Center Of Southeastern Wisconsin | PEN POLYMORPHISM OF HUMAN PLATELET MEMBRANE GLYCOPROTEIN IIIa AND DIAGNOSTIC AND THERAPEUTIC APPLICATION THEREOF |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4959308A (en) * | 1987-08-27 | 1990-09-25 | Board Of Regents, The University Of Texas System | Immunoassay for antibodies binding platelets |
US5110726A (en) * | 1987-08-27 | 1992-05-05 | Board Of Regents, The University Of Texas System | Immunoassay for antibodies binding platelets |
-
1989
- 1989-05-16 JP JP1122392A patent/JPH02300664A/en active Pending
-
1990
- 1990-04-12 US US07/508,922 patent/US5288610A/en not_active Expired - Lifetime
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4959308A (en) * | 1987-08-27 | 1990-09-25 | Board Of Regents, The University Of Texas System | Immunoassay for antibodies binding platelets |
US5110726A (en) * | 1987-08-27 | 1992-05-05 | Board Of Regents, The University Of Texas System | Immunoassay for antibodies binding platelets |
Non-Patent Citations (3)
Title |
---|
I. M. Roitt, Essential Immunology, 5th Edition, Blackwell Scientific Publications, Oxford, UK, 1984, pp. 159 161. * |
I. M. Roitt, Essential Immunology, 5th Edition, Blackwell Scientific Publications, Oxford, UK, 1984, pp. 159-161. |
W. J. Herbert, in D. M. Weir (Ed.), Handbook of Experimental Immunology, Third Edition, Blackwell Scientific Publications, Oxford, 1978, Chapter 20. * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0762121A1 (en) * | 1995-08-09 | 1997-03-12 | Nippon Zoki Pharmaceutical Co., Ltd. | Antigen for enzyme immunoassay and method of measuring anti-erythrocyte antibody |
Also Published As
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JPH02300664A (en) | 1990-12-12 |
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